Host Responses to Coccidioides by Human Airway Epithelium

人体气道上皮对球孢子菌的宿主反应

基本信息

批准号：
10373208
负责人：
Jatin M Vyas
金额：
$ 25.2万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-02 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10373208
关键词：
Address Animals Antifungal Antibiotics Apical Architecture Attenuated CRISPR screen Cell Communication Cell Culture Techniques Cell Line Cells Clustered Regularly Interspaced Short Palindromic Repeats Coccidioides Coccidioides immitis Coccidioides posadasii Complication Critical Pathways Data Direct Costs Disease Dissection Distal Environment Epithelial Epithelial Cells Extravasation Foundations Future Gene Expression Profile Genetic Transcription Host Defense Human Human body IL8 gene Immune Immune response Immunity Immunocompetent Immunocompromised Host Immunologics Individual Infection Inflammation Mediators Inflammatory Inhalation Innate Immune Response Investigation Knowledge Lead Lung Lung infections Mediating Methodology Methods Modeling Monitor Mus Mycoses Natural Resistance Pathogenicity Pathway interactions Patients Phagocytosis Play Proteins Response Elements Sampling Signal Pathway Signal Transduction Site Soil Sputum Surface System T cell response Tissues Transgenic Organisms United States Work airway epithelium base biosafety level 3 facility chemokine cytokine dimorphism experimental study fungus genome-wide in vivo innovation insight knockout gene lentiviral-mediated mortality mouse model new technology novel pathogen pathogenic fungus recruit response single-cell RNA sequencing stem cells transcriptome transcriptomics vaccine candidate

项目摘要

PROJECT SUMMARY Invasive fungal infections represent a major threat to immunocompromised patients and despite the availability of anti- fungal antibiotics, mortality rates remain as high as 50%. In the human host, the airway epithelium is the first point of contact upon inhalation of fungal conidia. As an immunologically active tissue, the airway epithelium may participate in active phagocytosis and coordinated immune cell recruitment. Soil fungi, including Coccidioides, do not need a human host to propagate, but upon disruption can enter the human body and lead to infection if not cleared. The fungal dimorph Coccidioides is native to arid regions in the southwestern U.S. and can establish infections in both immunocompetent and immunocompromised individuals. Unfortunately, little is known with regards to the initial response of human airway epithelium to Coccidioides. Currently, the Δcps1 strain of Coccidioides is being investigated as a potential vaccine candidate. Leveraging new technologies that permit the isolation of human airway stem cells, we demonstrated that primary differentiated human airway epithelial cells recapitulate the complexity of human airways. Our preliminary data showed infection at the apical surface of human airway epithelial cells by C. posadasii (Δcps1) elicits pro-inflammatory signals by the epithelium. Lastly, we demonstrated effective gene knockout in primary human airway epithelial cells using non-viral bulk nucleofection-based CRISPR strategy. Thus, our primary objective is to decipher the transcriptional and secretomal signatures critical to mount a successful response to Coccidioides in the human airway epithelium. To address our primary objective, we propose the following two specific aims: [1] determine the host response of human airway epithelium to both WT and the vaccine candidate C. posadasii (Δcps1), and [2] identify key signaling pathways to elicit an immune response to WT and the vaccine candidate C. posadasii (Δcps1) by human airway epithelium. This work will identify essential components of the human airway epithelium to mediate a protective response to these fungal infections and will provide the necessary foundation for future experiments to dissect the key pathways responsible for an effective host response to C. posadasii.

项目摘要侵入性真菌感染代表了对免疫功能低下的患者的主要威胁，并希望抗真菌抗生素，死亡率仍然高达50％。在人类主机中，气道上皮是第一点吸入真菌分生孢子后接触。作为一种免疫学性组织，气道上皮可能参与主动吞噬作用和协调的免疫细胞募集。土壤真菌，包括球虫，不需要人类宿主传播，但在干扰后可以进入人体并导致感染，如果未清除。真菌Dimorph 球虫剂原产于美国西南部干旱地区，可以在免疫能力和免疫功能低下的个体。不幸的是，关于人类气道的最初反应知之甚少上皮到球虫剂。目前，正在研究Coccidioides的ΔCPS1菌株作为潜在的疫苗候选人。利用允许人类气道干细胞隔离的新技术，我们证明了主要技术分化的人类气道上皮细胞概括了人类气道的复杂性。我们显示的初步数据 C. posadasii（ΔCPS1）在人气道上皮细胞的顶部表面感染，引起促炎信号上皮。最后，我们使用非病毒证明了原代人气道上皮细胞中有效的基因敲除基于块状核能的CRISPR策略。这是我们的主要目标是破译转录和秘密分解签名对于在人类气道上皮中成功地对球cid菌的成功反应至关重要。解决我们的初选目的，我们提出以下两个具体目标：[1]确定人类气道上皮对两者的宿主反应 WT和疫苗候选C. posadasii（ΔCPS1），[2]确定了关键信号通路以引起免疫反应人类气道上皮对WT和疫苗候选C. posadasii（ΔCPS1）。这项工作将确定重要的人类气道上皮的组成部分介导对这些真菌感染的受保护反应，并将提供将来实验的必要基础，以剖析负责有效宿主反应的关键途径。 posadasii。